US4702910A - Common antigen (PSC-A) to Pseudomonas aeruginosa which acts as an agent for pretecting Pseudomonas aeruginosa infection - Google Patents

Common antigen (PSC-A) to Pseudomonas aeruginosa which acts as an agent for pretecting Pseudomonas aeruginosa infection Download PDF

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US4702910A
US4702910A US06/797,796 US79779685A US4702910A US 4702910 A US4702910 A US 4702910A US 79779685 A US79779685 A US 79779685A US 4702910 A US4702910 A US 4702910A
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aeruginosa
psc
sup
antigen
mouse
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Tamotsu Fukuda
Shiro Shigeta
Hiroaki Okuya
Yasuyuki Kuroiwa
Tadashi Sudo
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Mitsui Toatsu Chemicals Inc
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Assigned to MITSUI TOATSU CHEMICALS, INC. reassignment MITSUI TOATSU CHEMICALS, INC. ASSIGNMENT OF ASSIGNORS INTEREST. Assignors: FUKUDA, TAMOTSU, KUROIWA, YASUYUKI, OKUYA, HIROAKI, SUDO, TADASHI, SHIGETA, SHIRO
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1203—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria
    • C07K16/1214—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-negative bacteria from Pseudomonadaceae (F)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/21—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Pseudomonadaceae (F)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
    • Y10S530/82—Proteins from microorganisms
    • Y10S530/825—Bacteria

Definitions

  • P. aeruginosa is originally known as an attenuated pathogenic bacterium.
  • the number of patients infected by P. aeruginosa as distinct from other bacterical infectious diseases has notably increased.
  • this disease After once occuring, the acute disease caused by this bacterum has a strong tendency to develop into a systemic infection. Also a good prognosis for this disease can not be expected. Therefore this disease has come to be numbered as one of the most difficient to cure cured bacterial infectious diseases.
  • This bacterium exhibits a high resistance to almost all of the antibiotics which have been typically used.
  • This bacterium tends to be resistant to recently developed antibiotics which are effective against P. aeruginosa.
  • aeruginosa corresponding to the monoclonal antibody produced by the C-Ab-producing hybridoma (hereafter referred to as C-Ab) by an affinity chromatography using immobilizing C-Ab.
  • C-Ab C-Ab-producing hybridoma
  • the common antigen obtained thus to P. aeruginosa which can react with C-Ab was found to be a completely new substance composed of protein, has ver low toxicity, and exhibits excellent ability in an animal experiment to protect infection by all serotypes of P. aeruginosa, resulting in this invention.
  • This invention also relates to an agent for protecting P. aeruginosa infection which contains the common antigen PSC-A as its active component.
  • the purpose of this invention is to supply an agent for protecting P. aeruginosa infection, which contains the common antigen PSC-A from P. aeruginosa as its active component for the vaccinotherapy of animals and humans.
  • strains of P. aeruginosa used in this invention are listed according to serological classification in Table 1. There are different theories about the classification of P. aeruginosa. The strains of P. aeruginosa used in this invention, are classified according to serological classification given in the report of the serotyping committee of the Japan P. aeruginosa society (1975, Japan J, Exp. Med., 46, 329, 1976). All strains belonging to A ⁇ M groups according to this classification can be used as a specific serotype of P. aeruginosa described in this invention. Since it was thought to be the best method, the classification of P. aeruginosa for this invention was performed according to the serological classifications established by the serotyping committee. Considering that new classification criteria will be adopted in the future, it can be said that the strains of P. aeruginosa which can be used in this invention, include all bacterial strains having PSC-A as an infection-protecting antigen.
  • C-Ab-producing hybridoma was produced according to the well-known method by Kohler, Milstein et al. (Nature, 256, 495, 1975). After an emulsion of P. aeruginosa ATCC 27581 (E type) treated with 0.3% of formalin was prepared with Freund incomplete adjuvant, this emulsion was intraperitoneally administered to female BALB/C mouse (7 weeks old) every other week for a total of 5 times to achieve immunization. The 5 ⁇ 10 3 mouse spleen cells collected 4 days following the final immunization and 5 ⁇ 10 7 NS-1 mouse myeloma cells were subjected to cell fusion in the presence of 50% polyethylene glycol to produce a hybridoma.
  • the hybridoma thus prepared was then poured into a 96-well flat bottom microplate and cultured on Dulbecco MEM medium supplemented with 10% fetal bovine serum containing HAT (hypoxanthine, amimopterin and thymidine) at 37° C. in the presence of 5% of CO 2 .
  • HAT hypoxanthine, amimopterin and thymidine
  • the presence of anit-P. aeruginosa monoclonal antibody in the culture solution was determined by Dot Immunobinding Assay which is an enzyme immunoassay (Anal. Biochem. 119, 142-147, 1982, hereafter referred to as DIBA method).
  • DIBA method was performed using a 96-well microtiter plate according to the following procedure.
  • a nitrocellulose membrane filter (3.1 mm square) prepared by immobilizing 0.4 ⁇ g per dot of P. aeruginosa treated with 0.3% of formalin (used as the antigen) was incubated with 100 ⁇ l of the above culture solution at room temperature for 30 minutes before being incubated with peroxidase-labelled anti-rabbit mouse immunoglobulin antibody (manufactured by DAKO company) for 30 minutes.
  • the nitrocellulose membrane filter was then reacted with 4-chloro-1-naphtol as a substrate for peroxidase. A positive result was recorded when dark blue spot was observed on the membrane filter by immobilizing the antigen.
  • the hybridoma was further subjected to cloning by limiting dilution.
  • the monoclonal hybridoma obtained thus was proliferation in a flask, and the proliferated monoclonal hybridoma was implanted in the abdominal cavity of a DALB/C mouse treated with the immunosuppressant Pristane (Aldrichi company).
  • Pristane immunosuppressant Pristane
  • ascites fluid of the treated mouse was applied to an affinity chromatography with Protein A-Sepharose (Pharmacia company) to prepare a purified monoclonal antibody.
  • the hybridomas obtained thus which can produce various anti-P.
  • aeruginosa monoclonal antibodies the inventors found a C-Ab-producing hybridoma which can produce a monoclonal antibody C-Ab reacting with all the serotypes of P. aeruginosa.
  • the reactivities of this C-Ab for several serotypes of P. aeruginosa determined by DIBA method are shown in Table 2.
  • This C-Ab exhibited almost equal affinities for all serotypes of P. aeruginosa. Additionally, this C-Ab did not react with either the endotoxin from serotype E of P. aeruginosa [lipopolysaccharied, prepared from the ATCC 27581 strain (N-10 strain) of P.
  • PSC-A Strains of P. aeruginosa which can be used in producing PSC-A are as described above. Conventional methods for culturing P. aeruginosa and crushing microbial cells may be used.
  • the medium heart infusion broth, brain heart infusion broth (manufactured by Eiken Kagaku), nutrient broth, or a synthetic medium prepared according to the method of Homma et al. (J. Biochem. 83, 711-18, 1978) can be used.
  • the synthetic medium by Homma et al. without any proteins is specially preferable because there is no possibility for contamination of proteins in the medium into microbial component. It is preferable that the temperature of the medium remain between 25° C.
  • the culture should be performed under aerobic conditions. For example, it is recommended that shaking culture or aerational agitation culture in a culture vessel should be performed.
  • the culture period influences the yield of PSC-A from P. aeruginosa.
  • a culture period of 16 ⁇ 24 hours is preferable.
  • the synthetic medium (pH7.4) of Homma et al. microbial cells desired were obtained by centrifugation of filtration.
  • the microbial cells obtained thus are then sufficiently mixed with water or a proper buffer solution before being crushed by a DYNOMILL, a French press, an ultrasonicator while being coolled below 10° C.
  • the suspension obtained thus is then decanted and subjected to centrifugation to obtain a cell-free extract of PSC-A.
  • the centrifugation of the suspension is performed at 39,000 ⁇ g for 30 minutes, and the supernatant is collected.
  • the rate for the extraction of PSC-A can be further improved by adding either a small quantity of a surfactant such as Triton X-100 a chelating agent such as EDTA or an enzyme such as lysozyme, deoxyribonuclease or ribonuclease.
  • a monoclonal antibody such as C-Ab with a specific affinity for PSC-A is used for the isolation and purification of PSC-A is used after immobilized with a proper carrier such as Affigel or CNBr-Sepharose as previously mentioned.
  • a proper carrier such as Affigel or CNBr-Sepharose as previously mentioned.
  • the affinity column to which the cell-free extract was applied is then thoroughly washed with a neutral buffer with a pH of 6 ⁇ 8 to elute contaminants other than PSC-A which are not bound to the above immobilized monoclonal antibody.
  • a buffer of low pH range usually used for the dissociation of the antigen-antibody complex such as 50 mM glycine-HCl buffered physiological saline (pH 3.0) is used for the affinity column to dissociate and elute PSC-A bound to the monoclonal antibody.
  • the eluate prepared thus is then adjusted pH to be neutral before the neutralized solution is dialyzed against distilled water. After that, the dialysate is lyophilized to obtain a pure powder of PSC-A.
  • the concentration of PSC-A in the filtrate from the culture can be neglected as long as microbial cells are obtained as a usual culture product of living bacteria.
  • a large amount of PSC-A has been transferred from microbial cells to a filtrate due to death or autolysis of microbial cells by a long period culture, it is possible to recover the purified PSC-A by applying a supernatant by centrifugation of the above filtrate to an affinity column prepared by immobilizing the above monoclonal antibody with a specific affinity for PSC-A.
  • Protein Content (%) : 55.0 [(colorimetric analysis by hydrolysis using ninhydrin, bovine serum albumin as the standard)(Anal. Biochem., 49, 95, 1972)] 27.0 [modified Lowry method, bovine serum albumin as the standard) (Anal. Biochem., 69, 646, 1975)], 24.0 [(protein binding assay, bovine serum albumin as the standard)(Anal. Biochem., 72, 248, 1976)].
  • Hexosamine Content (%) : below 1.0 [(Rondle. Morgan method, glucosamine as the standard)(Biochem. J., 61, 586, 1955)].
  • the original spot is colored when the chloroform methanol extract of PSC-A (modified Bligh. Dyer method) is developed on a silica gel TLC by using a mixture of petroleum ether, ether and ocetic acid (80:20:1) before being colored by treating with 50 % sulfuric acid solution and heating it.
  • PSC-A modified Bligh. Dyer method
  • the mouse monoclonal antibody having a specific affinity for PSC-A does not react with either the well-known common antigen OEP to P. aeruginosa or the endotoxin of P. aeruginosa (lipopolysaccharide).
  • this agent When used as an agent for protecting against P. aeruginosa infection, it is preferable that this agent be administered by injection.
  • a solution or a lyophilized preparation prepared from PSC-A alone or combining it with usual additive and excipient can be practically used. It is possible to incorporate PSC-A in an oil-in-water type emulsion or a water in-oil type emulsion. Also, PSC-A can be practically used either by sealing PSC-A in liposome composed of phospholipid, cholesterol or by fixing PSC-A to the outer surface of the membrane of liposome.
  • the dose and the administration route for PSC-A may be properly selected. It is preferable that the dose be 0.001 ⁇ 10 mg per kg body weight.
  • intracutaneous, subcutaneous, intravenous, intramuscular and intraperitoneal administrations can be performed.
  • PSC-A has antigenicity which occurs in a so-called vaccine and induces the production of high concentration of an antibody for P. aeruginosa in the serum of a mouse or guinea pig immunized with PSC-A.
  • PSC-A is highly active for protecting animal P. aeruginosa infection.
  • mice immunized with PSC-A were not infected with a lethal does of each of all serotypes of P. aeruginosa and could live after being treated with P. aeruginosa. This indicates remarkable ability of PSC-A protect P. aeruginosa infection.
  • PSC-A has low acute toxicity because the 50% lethal does of PSC-A when it is intravenously administered to mice is more than 5 mg/kg. As shown in claim, PSC-A does not cause any direct toxicity in animal cells.
  • PSC-A is very useful as an agent for protecting against P. aeruginosa infection.
  • the medium for the culture of P. aeruginosa was contained (per liter) 20 g of sodium glutamate, 5 g of glycerin, 0.1 g of MgSO 4 .7H 2 O, 0.55 g of KH 2 PO 4 , 5.6 g of Na 2 HPO 4 .12H 2 O, 17.26 mg of Ca(NO 3 ) 2 .4H 2 O and 50 ⁇ g of FeSO 4 .7H 2 O, and was adjusted to pH 7.6.
  • P. aeruginosa ATCC27581 (E type) was cultured on a nutrient agar medium at 37° C. overnight, microbial cells were suspended in physiological saline.
  • The, 0.5 ml of the suspension was inoculated into a Sakaguchi's flask containing 150 ml of the synthetic medium having the above composition, and shaking culture was performed at 37° C. for 16 hours.
  • the wet microbial cells obtained thus were then suspended in 220 ml of 20 mM Tris-Hc buffer (pH 8.0) containing 2% of Triton X 100 and 10 mM of EDTA before being crushed with a DYNO-MILL (beads 0.1 mm ⁇ ) for three minutes while being cooled.
  • the column was then washed with 100 ml of 20 mM Tris-HCl buffer (pH 8.0) containing 0.5% Triton X100 and 100 ml of 20 mM Tris-HCl buffer (pH 8.0), respectively and eluted with 60 mlof 50 mM glycine-HCl buffered saline (pH 3.0).
  • the eluate obtained thus was then neutralized with 1N aqueous sodium hydrogen carbonate solution and dialyzed against distilled water at 4° C. for 24 hours. After that, the dialyzate was lyophilized to obtain 4.7 mg of a pure powder of PSC-A.
  • P. aeruginosa ATCC 27584 (G type) was subjected to shaking culture in the synthetic medium (in the same way as in Example 1) at 37° C. for 20 hours. The below procedure was performed in the same way as in Example 1. 6.2 mg of PSC-A was obtained from 58.2 g of wet microbial cells.
  • the amino acid composition of PSC-A was determined by an automatic amino acid analyzer (IRICA A-5500) and the results are shown in Table 3.
  • P. aeruginosa ATCC 27586 (I type) was subjected to shaking culture in the synthetic medium (in the same way as in Example 1) at 37° C. for 20 hours. The below procedure was performed in the same way as in Example 1. 28 mg of PSC-A was obtained from 305 g of wet microbial cells.
  • Example 2 After 1 mg of PSC-A obtained in Example 2 was dissolved in 10 ml of physiological saline, the solution prepared thus was subjected to aseptic filtration through a Nuclepore NO 20 (manufactured by Nuclepore company). A 1 ml sample of the filtrate obtained thus was then poured into each vial without allowing any contamination by bacteria to obtain a solution of PSC-A.
  • the solution was filtered through a Nuclepore NO 20. A 1 ml sample of the filtrate obtained thus was then poured into each vial without allowing any contamination by bacteria before lyophilization was performed to obtain a lyophilized preparation of PSC-A.
  • Example 3 After 1 mg of PSC-A obtained in Example 3 was dissolved in 0.5 ml of physiological saline, 0.5 ml of mixture solution consisting of liquid paraffin and Arlacel (manufactured by Arlacel A. Atlas Chemical Industries) in a ratio of 8.5 to 1.5 was added to the solution and a water-in-oil type emulsion was prepared.
  • mixture solution consisting of liquid paraffin and Arlacel (manufactured by Arlacel A. Atlas Chemical Industries) in a ratio of 8.5 to 1.5 was added to the solution and a water-in-oil type emulsion was prepared.
  • Equal amounts of Freund incomplete adjuvant and a physiological saline solution of PSC-A obtained in Example 1 and that in Example 2 are mixed to prepare water-in-oil type emulsions, respectively.
  • mice After two groups each consisting of BALB/C female mice, 8 weeks old were immunized twice with each of the two emulsions shown in Experimental Example 2 at one-week intervals (10 ⁇ g per mouse of PSC-A was intraperitoneally administered for each immunization), each mouse was infected with P. aeruginosa (5 LD 50 ) one week following the final immunization. Two strains of P. aeruginosa PA103 (E type) and P 28 (C type) were used for the infection. Each of these strains was cultured on a heart infusion agar medium (manufactured by Eiken Kagaku) overnight and collected and diluted with physiological saline. The 5 pts.
  • P. aeruginosa PA103 E type
  • P 28 C type
  • PSC-A obtained in Example 3 was dissolved in physiological saline to prepare a sample solution. After two groups each consisting of five BALB/C female mice, 8 weeks old were immunized four times with the solution prepared thus at one-week intervals (20 ⁇ g per mouse of PSC-A was subcutaneously administered for each immunization), these two groups were infected with two strains of P. aeruginosa consisting of Pa103 (E type) and P28 (G type), respectively. A bacterial suspension of each of these strains was prepared in the same manner as in Example 3 and each mouse was infected with the bacterium by being intraperitoneally inoculated with an about 3 LD 50 amount of the bacterial suspension. For the control group, physiological saline alone was administered instead of PSC-A.
  • Example 1 Each of PSC-A obtained in Example 1 and that in Example 2 was dissolved in physiological saline to prepare sample solutions.
  • Two groups each consisting of 15 female mice, 6 weeks old were immunized three times with each of the two sample solutions at one-week intervals (30 ⁇ g per mouse of PSC-A was intraperitoneally administered for each immunization).
  • One week following the final immunization each mouse was infected with P. aeruginosa.
  • Two strains of P. aeruginosa PA103 (E type) and P28 (G type) were used for the infection similarly to Experimental Example 3 and 4.

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US06/797,796 1984-05-25 1985-11-14 Common antigen (PSC-A) to Pseudomonas aeruginosa which acts as an agent for pretecting Pseudomonas aeruginosa infection Expired - Fee Related US4702910A (en)

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FR (1) FR2590172B1 (enrdf_load_stackoverflow)
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4946677A (en) * 1986-06-24 1990-08-07 Immuno Aktiengesellschaft Fur Chemisch-Medizinische Produkte Preparations active against pseudomonas aeruginosa infections and methods of producing them
US5114712A (en) * 1985-11-14 1992-05-19 Mitsui Toatsu Chemicals, Inc. Common antigen (PSC-A) to Pseudomonas aeruginosa which acts as an agent for protecting Pseudomonas aeruginosa infection
US5179001A (en) * 1985-09-27 1993-01-12 The Regents Of The University Of California Detection and monitoring of chronic and gram-negative infection
US5237053A (en) * 1986-06-24 1993-08-17 Immuno Aktiengesellschaft Preparation active against Pseudomonas aeruginosa infections and methods of producing them
US5662905A (en) * 1985-12-10 1997-09-02 Bristol-Myers Squibb Company Monoclonal antibody compositions cross-reactive and cross-protective against P. aeruginosa serotypes
RU2155226C2 (ru) * 1993-06-07 2000-08-27 Чейл Фудз энд Кемикалз, Инк. Новый ослабленный штамм pseudomonas aeruginosa

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AT390192B (de) * 1988-08-29 1990-03-26 Immuno Ag Gegen pseudomonas aeruginosa-infektionen wirksame praeparationen sowie immunglobuling-h|ltige, gegen bakterium pseudomonas aeruginosa wirksame praeparationen
JPH0550792A (ja) * 1991-08-22 1993-03-02 Oohira:Kk ノツクシヤープペンシルにおける芯ロツク機構
DE4128454A1 (de) * 1991-08-28 1993-03-04 Riedel De Haen Ag Monoklonaler antikoerper, der mit pseudomonas aeruginosa reagiert
JPWO2007049770A1 (ja) * 2005-10-28 2009-04-30 明治製菓株式会社 緑膿菌の外膜タンパク質ｐａ５１５８
JP2009132686A (ja) * 2007-10-26 2009-06-18 Okayama Univ ボツリヌス毒素由来のポリペプチド及びボツリヌス毒素の検出方法
CN108508743B (zh) * 2018-06-25 2021-06-01 长沙理工大学 时滞系统的准pi预测控制新方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2290219A1 (fr) * 1974-11-05 1976-06-04 Pasteur Institut Principe actif de vaccin antipyocyanique
US4428931A (en) * 1982-03-15 1984-01-31 Merck & Co., Inc. Bacterial toxoids and gram-negative immune globulin therefrom
US4575459A (en) * 1984-02-24 1986-03-11 Toho Yakuhin Kogyo Kabushiki Kaisha Toxoids of elastase of Pseudomonas aeruginosa origin
US4578458A (en) * 1983-03-23 1986-03-25 Brigham And Women's Hospital Mucoid exopolysaccharide vaccine against Pseudomonas aeruginosa

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2460139A1 (fr) * 1979-06-29 1981-01-23 Pasteur Institut Fraction antigenique glycopeptidique vaccinante a tres grande immunogenicite isolee de cultures de germes pathogenes, procedes d'isolement de cette fraction et vaccins contenant ladite fraction

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2290219A1 (fr) * 1974-11-05 1976-06-04 Pasteur Institut Principe actif de vaccin antipyocyanique
US4428931A (en) * 1982-03-15 1984-01-31 Merck & Co., Inc. Bacterial toxoids and gram-negative immune globulin therefrom
US4578458A (en) * 1983-03-23 1986-03-25 Brigham And Women's Hospital Mucoid exopolysaccharide vaccine against Pseudomonas aeruginosa
US4575459A (en) * 1984-02-24 1986-03-11 Toho Yakuhin Kogyo Kabushiki Kaisha Toxoids of elastase of Pseudomonas aeruginosa origin

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5179001A (en) * 1985-09-27 1993-01-12 The Regents Of The University Of California Detection and monitoring of chronic and gram-negative infection
US5114712A (en) * 1985-11-14 1992-05-19 Mitsui Toatsu Chemicals, Inc. Common antigen (PSC-A) to Pseudomonas aeruginosa which acts as an agent for protecting Pseudomonas aeruginosa infection
US5662905A (en) * 1985-12-10 1997-09-02 Bristol-Myers Squibb Company Monoclonal antibody compositions cross-reactive and cross-protective against P. aeruginosa serotypes
US4946677A (en) * 1986-06-24 1990-08-07 Immuno Aktiengesellschaft Fur Chemisch-Medizinische Produkte Preparations active against pseudomonas aeruginosa infections and methods of producing them
US5237053A (en) * 1986-06-24 1993-08-17 Immuno Aktiengesellschaft Preparation active against Pseudomonas aeruginosa infections and methods of producing them
RU2155226C2 (ru) * 1993-06-07 2000-08-27 Чейл Фудз энд Кемикалз, Инк. Новый ослабленный штамм pseudomonas aeruginosa

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GB8528191D0 (en) 1985-12-18
JPH0526474B2 (enrdf_load_stackoverflow) 1993-04-16
FR2590172B1 (fr) 1987-12-24
JPS60248625A (ja) 1985-12-09
FR2590172A1 (fr) 1987-05-22
DE3541044A1 (de) 1987-05-21
GB2182937A (en) 1987-05-28
GB2182937B (en) 1989-11-01

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